富氢水联合丙泊酚后处理对乳鼠离体脑片谷氨酸损伤后保护作用的研究

发布时间：2018-03-11 09:20

本文选题：富氢水　切入点：丙泊酚　出处：《贵阳医学院》2015年硕士论文　论文类型：学位论文

【摘要】：目的:1、研究富氢水对乳鼠离体脑片谷氨酸损伤后的保护作用,并探讨其在乳鼠离体脑片损伤后保护作用的最佳浓度;2、研究富氢水(hydrogen-rich water)联合丙泊酚(propofol)后处理对谷氨酸(L-Glutamic Acid,Glu)致乳鼠离体脑片损伤的保护作用,并探讨其机制。方法:1、切取并培养出生7天的SD大鼠乳鼠离体脑片,随机分为10个小组。实验分两个步骤进行。第一步:探索富氢水最佳保护浓度,5个小组分别为:25μmol/l富氢水+谷氨酸损伤组(HL25+Glu组)、50μmol/l富氢水+谷氨酸损伤组(HL50+Glu组)、100μmol/l富氢水+谷氨酸损伤组(HL100+Glu组)、200μmol/l富氢水+谷氨酸损伤组(HL200+Glu组)、300μmol/l富氢水+谷氨酸损伤组(HL300+Glu组);第二步,探讨富氢水联合丙泊酚的保护作用,5个小组分别为:正常对照组,Glu损伤组(Glu损伤组),3 mg/L丙泊酚处理组(PL3+Glu处理组),100μmol/l富氢水后处理组(HL100+Glu处理组),富氢水丙泊酚联合组(HL100+PL3+Glu处理组)。每组12例。将Glu损伤组、HL25+Glu组、HL50+Glu组、HL100+Glu组、HL200+Glu组、HL300+Glu组及PL3+Glu、HL100+Glu、HL100+PL3+Glu处理组均培养至第六天,用含1 mmol/L Glu的培养基损伤30min,建立缺血再灌注模型,损伤后将Glu损伤组移入正常的完全培养基,将HL25+Glu组、HL50+Glu组、HL100+Glu组、HL200+Glu组、HL300+Glu组及PL3+Glu、HL100+Glu、HL100+PL3+Glu处理组分别移入含相应处理药物的完全培养基中继续培养,每3小时换液一次,共8次,24小时。不同富氢水浓度组检测尼氏小体数量,药物处理组检测苏木素一伊红(HE)染色,尼氏小体计数、乳酸脱氢酶(Lactate dehydrogenase LDH)释放率、碱性成纤维生长因子(b FGF)数目的测定来比较各组脑片神经细胞的损伤程度。结果:1、与富氢水其他浓度组相比,浓度为100μmol/l的富氢水后处理组的尼氏小体数量最多;2、与正常对照组相比,Glu损伤后的各组脑片中神经细胞均受到一定程度的损伤,表现为细胞数量减少,细胞膜破碎,胞核皱缩,LDH释放率、b FGF数量明显增高;尼氏小体数量减少;3、与Glu损伤组相比,富氢水及丙泊酚后处理的各组,均能改善脑片中神经细胞形态结构受损的情况,细胞膜破损减轻,胞核较完整,均能使尼氏小体数量增加,LDH释放率及b FGF数量降低,而两者之间差异均无统计学意义;4、富氢水联合丙泊酚后处理组的尼氏小体数量较丙泊酚药物后处理组增加,LDH释放率及b FGF数量均降低;与100μmol/l的富氢水后处理组相较,药物联合组的尼氏小体数量增加,而LDH释放率及b FGF数量均增加。结论:1、不同浓度的富氢水对谷氨酸介导的乳鼠离体脑片损伤确有一定的保护作用,其中以100μmol/l为最佳;2、丙泊酚对乳鼠离体脑片谷氨酸损伤后的保护作用确切;3、富氢水联合丙泊酚对缺血再灌注损伤的保护作用较单独使用效果更好,其机制可能与减轻神经胶质细胞过度活化有关。
[Abstract]:Objective to study the protective effect of hydrogen rich water on glutamate injury in isolated brain slices of neonatal rats. To study the protective effect of propofol propofolate combined with hydro-rich water on the injury induced by L-Glutamic Acidic acid (GluA) in isolated rat brain slices, and to investigate the protective effect of the optimal concentration of Glutamic acid on the injury of isolated rat brain slices induced by hydrogen rich water and propofol (P < 0.05). Methods the isolated brain slices of SD rats born 7 days old were cut and cultured at 1: 1. The experiment was divided into 10 groups randomly. The experiment was divided into two steps. The first step was to explore the best protection concentration of hydrogen rich water. Five groups were divided into 5 groups: 1: 25 渭 mol/l hydrogen rich water glutamate injury group, HL25 Glu group, 50 渭 mol/l hydrogen rich water glutamate injury group, HL-50 Glu group, 100 渭 mol/l. HL-100 Glu group, HL-200 Glu group, HL-200 Glu group, HL-300 Glu group, HL-300 Glu group, HL-200 Glu group, HL-300 Glu group, HL-100 Glu group, HL-200 Glu group, HL-200 Glu group, HL-300 Glu group, HL-200 Glu group, HL-200 Glu group, HL-300 Glu group. To investigate the protective effect of hydrogen rich water combined with propofol, the five groups were: normal control group, Glu injury group, mg/L injury group, propofol treatment group, mg/L Glu treatment group, 100 渭 mol/l hydrogen rich water post-treatment group, HL-100 Glu treatment group, hydrogen rich water propofol group, and hydrogen rich water propofol group. 12 cases in each group were treated with HL-100 PL3 Glu. All of them were cultured to 6th days in Glu injury group, HL-25 Glu group, HL50 Glu group, HL100 Glu group, HL200 Glu group, HL300 Glu group and PL3 Glull-HL100 Glu-HL100 PL3 Glu group. The model of ischemia-reperfusion was established by using the medium containing 1 mmol/L Glu for 30 minutes. After the injury, the Glu injury group was transferred into the normal complete medium. HL25 Glu group, HL-50 Glu group, HL-100 Glu group, HL-200 Glu group, HL-300 Glu group and PL3 GluH100 Glutin-HL100 PL3 Glu group were cultured in complete culture medium containing corresponding drugs respectively, and the solution was changed every 3 hours. The number of Nissl bodies was detected in different concentrations of hydrogen rich water, the staining of hematoxylin-eosin (HEH), the count of Nissl bodies and the release rate of lactate dehydrogenase (Lactate dehydrogenase LDHs) were detected in the drug treatment group. The number of basic fibroblast growth factor (bFGFs) was measured to compare the degree of nerve cell damage in brain slices in each group. Results: 1, compared with other concentrations of hydrogen rich water, Compared with the normal control group, the number of Nissl corpuscles in the 100 渭 mol/l hydrogen-rich water treatment group was the highest, and the nerve cells in the brain slices were damaged to a certain extent, which showed that the number of cells decreased and the cell membrane was broken, compared with the normal control group. The release rate of LDH and the number of Nissl corpuscles were significantly increased, and the number of Nissl bodies was decreased. Compared with the Glu injury group, all groups treated with hydrogen rich water and propofol could improve the damage of neuronal morphology and cell membrane in brain slices, and the damage of cell membrane was alleviated in all the groups treated with hydrogen rich water and propofol. Nuclear integrity increased the release rate of FGF and the number of b FGF in Nissl corpuscles. However, there was no significant difference between the two groups. The number of Nissl bodies in the hydrogen-rich water combined with propofol post-treatment group increased the release rate of FGF and the number of b FGF in the propofol post-treatment group, which was significantly lower than that in the post-treatment group with 100 渭 mol/l hydrogen rich water. In the drug combination group, the number of Nissl bodies increased, while the release rate of LDH and the number of b FGF increased. Conclusion different concentrations of hydrogen rich water have protective effect on glutamate mediated brain injury in isolated rat brain slices. The protective effect of propofol on glutamate injury in isolated brain slices of neonatal rats was 100 渭 mol/l. The protective effect of hydrogen rich water combined with propofol on ischemia-reperfusion injury was better than that of using propofol alone, and the protective effect of propofol on ischemia reperfusion injury was better than that of propofol alone. The mechanism may be related to reducing the excessive activation of glial cells.
【学位授予单位】：贵阳医学院
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R614

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